RNA immunoprecipitation to study RNA-protein interactions (CAT#: STEM-MB-0163-WXH)

Introduction

RNA immunoprecipitation (RIP) mainly uses antibodies against the target protein to precipitate the corresponding RNA-protein complex, and after separation and purification, the RNA bound to the complex can be detected by q-PCR or sequencing. RIP is a technique for studying the combination of RNA and protein in cells. It is a powerful tool for understanding the dynamic process of post-transcriptional regulatory network and can help us discover the regulatory targets of miRNA.

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Applications Procedure

Applications

• Study of RNA and protein binding in cells.
• Discovery of regulatory targets of miRNAs.

Procedure

1. Harvest cells
2. Isolate the nucleus and lyse the nucleus pellet
3. Chromatin Fragmentation
4. Immunoprecipitation of the RNA-binding protein (RBP) of interest along with the bound RNA
5. Wash away unbound material
6. Purification of Bound RNA on RBP After Immunoprecipitation
7. Reverse Transcription of RNA to cDNA for Analysis by qPCR, Microarray, or Sequencing

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